title: miR-Control
description: Protocols: MP41 cell line was obtained from Decaudin's lab at Curie Institute, Paris, France. Mel202 and 92.1 cell line were obtained from European Collection of Authenticated Cell Cultures (ECACC) (Merck). All cell lines were maintained in humidified air (37°C, 5% CO2). Uveal melanoma cell lines were maintained in RPMI-1640 medium (GibcoTM, ThermoFisher, MA) supplemented with 20% Fetal Bovine Serum (FBS) (EurobioScientific) and 1% Penicillin-Streptomycin (PS) antibiotics (GibcoTM, ThermoFisher). All cell lines have been routinely tested for mycoplasma contamination (Mycoplasma contamination detection kit; rep-pt1; InvivoGen). Experiments were performed on MP41 cells according to the protocol published by Judy Lieberman's lab (Tan SM, Lieberman J. Capture and identification of mirna targets by biotin pulldown and RNA-seq. In: Methods in Molecular Biology. 2016. p. 211-28.) but with minor modifications. 15 millions of MP41 cells were seeded in 3 X 150mm dishes 5 millions each) and were transfected the next day with 100nM of biotinylated miR-16 or miR-CTR (Dharmacon). Next, they were harvested ~24h post transfection. Cells from the 3 dishes were treated separately. Meanwhile, magnetic beads (Streptavidine Dynabeads™ M-280 DYNAL™, ThermoFisher Scientific) were washed and blocked according to the protocol. Cell lysate and washed beads were incubated for 4 hours at 4°C on a rotating agitator. All next steps: the precipitation and the purification of coupled RNA was performed according to the protocol. We pooled the 3 same conditions (from the 3 transfected plates) in the end of the RNA purification. Purified RNA was quantified using quantified using a NanoDrop 1000 Spectrophotometer (ThermoFisher Scientific) Experiments were performed on MP41 cells according to the protocol published by Judy Lieberman's lab (Tan SM, Lieberman J. Capture and identification of mirna targets by biotin pulldown and RNA-seq. In: Methods in Molecular Biology. 2016. p. 211-28.).
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2022-09-01
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2022-09-01
attributes.tag: External Id
attributes.value: SAMEA10110056
attributes.tag: INSDC center alias
attributes.value: INSERM U1242 OSS (Oncogenesis, Stress and Signaling) - Universite de Rennes1
attributes.tag: INSDC center name
attributes.value: INSERM U1242 OSS (Oncogenesis, Stress and Signaling) - Universite de Rennes1
attributes.tag: INSDC first public
attributes.value: 2022-09-01T00:13:45Z
attributes.tag: INSDC last update
attributes.value: 2022-09-01T00:13:45Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-10940:miR-Control
attributes.tag: age
attributes.value: 49
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell_line
attributes.value: MP41
attributes.tag: common name
attributes.value: human
attributes.tag: developmental stage
attributes.value: adult
attributes.tag: disease
attributes.value: uveal melanoma
attributes.tag: organism part
attributes.value: eye
attributes.tag: sample name
attributes.value: E-MTAB-10940:miR-Control
attributes.tag: sex
attributes.value: female
attributes.tag: treatment
attributes.value: biotinylated miR-Control transfection